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MicroRNA-939 down-regulates CD2-associated protein by targeting promoter in HEK-293T cells

Yu-Ping Huang1, Ling-Zhi Qiu2, Guo-Ping Zhou1

  • 1a Department of Pediatrics , the First Affiliated Hospital, Nanjing Medical University , Nanjing , Jiangsu , China ;

Renal Failure
|February 20, 2016
PubMed

Insights

MicroRNAs (miRNAs) can suppress CD2-associated protein (CD2AP) expression transcriptionally. Specifically, miR-939 targets CD2AP promoter sequences, reducing CD2AP levels and potentially contributing to podocyte injury.

Area of Science:

  • Molecular biology
  • Genetics
  • Nephrology

Background:

  • CD2-associated protein (CD2AP) is crucial for maintaining podocyte integrity and preventing proteinuria.
  • MicroRNAs (miRNAs) are key regulators of gene expression, and their loss in podocytes can cause significant proteinuria.

Purpose of the Study:

  • To investigate the role of miRNAs in regulating CD2AP expression at the transcriptional level.
  • To identify specific miRNAs that target and suppress CD2AP gene expression.

Main Methods:

  • In silico analysis of the human CD2AP promoter for miRNA binding sites.
  • Transfection of candidate miRNAs (miR-939, miR-148b*, miR-191*, miR-638) into HEK-293T cells.
  • Dual-luciferase reporter assays to assess promoter activity.
  • Analysis of mRNA and protein levels of CD2AP.

Main Results:

  • miR-939 was identified as a miRNA that significantly reduces CD2AP promoter activity.
  • miR-939 was confirmed to down-regulate both mRNA and protein expression of CD2AP.
  • The study identified a transcriptional regulatory mechanism for CD2AP by miR-939.

Conclusions:

  • miR-939 directly targets sequences within the CD2AP promoter, suppressing its gene expression.
  • These findings suggest a novel mechanism by which miRNAs can contribute to podocyte injury by reducing essential slit diaphragm proteins like CD2AP.
Keywords:
CD2-associated proteingene expression regulationmicroRNA-939podocyte injurypromoter

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